3D virtual histology of human pancreatic tissue by multiscale phase-contrast X-ray tomography.

Jasper Frohn,Diana Pinkert-Leetsch,Tim Salditt,Frauke Alves,Marius Reichardt,Markus Osterhoff,Jeannine Missbach-Güntner

doi:10.1107/s1600577520011327

Abstract

A multiscale three-dimensional (3D) virtual histology approach is presented, based on two configurations of propagation phase-contrast X-ray tomography, which have been implemented in close proximity at the GINIX endstation at the beamline P10/PETRA III (DESY, Hamburg, Germany). This enables the 3D reconstruction of characteristic morphological features of human pancreatic normal and tumor tissue, as obtained from cancer surgery, first in the form of a large-scale overview by parallel-beam illumination, followed by a zoom into a region-of-interest based on zoom tomography using a Kirkpatrick-Baez mirror with additional waveguide optics. To this end 1 mm punch biopsies of the tissue were taken. In the parallel tomography, a volumetric throughput on the order of 0.01 mm3 s-1 was achieved, while maintaining the ability to segment isolated cells. With a continuous rotation during the scan, a total acquisition time of less than 2 min was required for a full tomographic scan. Using the combination of both setups, islets of Langerhans, a three-dimensional cluster of cells in the endocrine part of the pancreas, could be located. Cells in such an islet were segmented and visualized in 3D. Further, morphological alterations of tumorous tissue of the pancreas were characterized. To this end, the anisotropy parameter Ω, based on intensity gradients, was used in order to quantify the presence of collagen fibers within the entire biopsy specimen. This proof-of-concept experiment of the multiscale approach on human pancreatic tissue paves the way for future 3D virtual pathology.

Highlights

This enables the 3D reconstruction of characteristic morphological features of human pancreatic normal and tumor tissue, as obtained from cancer surgery, first in the form of a large-scale overview by parallel-beam illumination, followed by a zoom into a region-of-interest based on zoom tomography using a Kirkpatrick–Baez mirror with additional waveguide optics
In this work we have performed multiscale 3D virtual histology of human pancreatic biopsies combining two configurations of X-ray propagation imaging with synchrotron radiation
Since cancer surgery is often confronted with removal of substantial tissue volumes, 3D patho-histology automatically faces a multiscale challenge

Summary

Introduction

The potential of propagation-based X-ray phase-contrast tomography for three-dimensional (3D) virtual histology has been pointed out by a number of recent studies (Saccomano et al, 2018; Topperwien et al, 2018; Khimchenko et al, 2016; Albers et al, 2018; Dejea et al, 2019; Ding et al, 2019; De Clercq et al, 2019; Strotton et al, 2018; Mei et al, 2020). 3D virtual histology by phase-contrast X-ray tomography is able to extend classical histological two-dimensional (2D) approaches, which are based on thin sections and light microscopy, into a full 3D visualization with isotropic resolution and without destructive slicing of the specimen. Compared with other full-field phase-contrast techniques, e.g. based on conventional grating interferometry or analyzer crystals, propagation-based imaging (PBI) can reach a resolution below optical microscopy (Khimchenko et al, 2018)

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